Samuelson J C, Caulfield J P, David J R
J Cell Biol. 1982 Aug;94(2):355-62. doi: 10.1083/jcb.94.2.355.
The lectin concanavalin A (Con A) was used as a model probe to study the behavior of molecules bound to the surface of recently transformed schistosomula of Schistosoma mansoni. Con A binding was saturable (150-180 pg/organism) and specifically competed by alpha-methyl mannoside. Both FITC-Con A and 125-I-Con A were lost from the surface of schistosomula with a halftime of 8-10 h in culture in defined medium. A comparable decrease in the binding of Con A to schistosomula cultured and then labeled with the lectin indicated that the labeling procedure itself was not inducing the observed change. Internalization of Con A was not seen by either fluorescence microscopy or electron microscope radioautography. In addition, 70-80% of the radioactivity lost from the parasite was recoverable by TCA precipitation from the culture medium as intact Con A (27,000 mol wt on SDS PAGE). Thus, the mechanism of clearance of bound Con A from the surface of cultured schistosomula is apparently by sloughing of Con A molecules intact into the culture media and not by endocytosis and degradation. Con A binding sites, visualized with hemocyanin by scanning electron microscopy, appeared homogeneously distributed over the surface of schistosomula when organisms were labeled at 4 degree C or after fixation with glutaraldehyde. However, Con A and hemocyanin formed aggregates on the surface of schistosomula when labeling was performed at 37 degrees C, which suggests that lectin binding sites have lateral mobility within the plane of the membrane. These aggregates are likely independent of metabolism by the parasite because aggregation also occurs on the surface of organisms killed with azide.
刀豆球蛋白A(Con A)被用作模型探针,以研究与曼氏血吸虫新近转化的童虫表面结合的分子行为。Con A的结合是可饱和的(150 - 180 pg/虫体),并可被α-甲基甘露糖苷特异性竞争。在限定培养基中培养时,FITC-Con A和¹²⁵I-Con A均以8 - 10小时的半衰期从童虫表面丢失。用Con A培养然后标记的童虫,其Con A结合的类似减少表明标记过程本身并未诱导观察到的变化。通过荧光显微镜或电子显微镜放射自显影均未观察到Con A的内化。此外,从寄生虫中丢失的70 - 80%的放射性可通过三氯乙酸沉淀从培养基中回收为完整的Con A(SDS-PAGE上分子量为27,000)。因此,从培养的童虫表面清除结合的Con A的机制显然是Con A分子完整地脱落到培养基中,而不是通过内吞作用和降解。当在4℃标记或用戊二醛固定后,用扫描电子显微镜观察到的与血蓝蛋白结合的Con A位点似乎均匀分布在童虫表面。然而,当在37℃进行标记时,Con A和血蓝蛋白在童虫表面形成聚集体,这表明凝集素结合位点在膜平面内具有侧向流动性。这些聚集体可能与寄生虫的代谢无关,因为在叠氮化物杀死的虫体表面也会发生聚集。